Version 1
: Received: 6 July 2020 / Approved: 7 July 2020 / Online: 7 July 2020 (09:49:17 CEST)
How to cite:
Jo, S.; Jang, E.; Kim, M.; Kim, S.; Lee, K.; Shin, D.H. A Tryptophan-based Assay Method to Search Regulatory Compounds for TCTP. Preprints2020, 2020070122. https://doi.org/10.20944/preprints202007.0122.v1
Jo, S.; Jang, E.; Kim, M.; Kim, S.; Lee, K.; Shin, D.H. A Tryptophan-based Assay Method to Search Regulatory Compounds for TCTP. Preprints 2020, 2020070122. https://doi.org/10.20944/preprints202007.0122.v1
Jo, S.; Jang, E.; Kim, M.; Kim, S.; Lee, K.; Shin, D.H. A Tryptophan-based Assay Method to Search Regulatory Compounds for TCTP. Preprints2020, 2020070122. https://doi.org/10.20944/preprints202007.0122.v1
APA Style
Jo, S., Jang, E., Kim, M., Kim, S., Lee, K., & Shin, D.H. (2020). A Tryptophan-based Assay Method to Search Regulatory Compounds for TCTP. Preprints. https://doi.org/10.20944/preprints202007.0122.v1
Chicago/Turabian Style
Jo, S., Kyunglim Lee and Dong Hae Shin. 2020 "A Tryptophan-based Assay Method to Search Regulatory Compounds for TCTP" Preprints. https://doi.org/10.20944/preprints202007.0122.v1
Abstract
Transcriptionally controlled tumor protein (TCTP) is a highly conserved protein performing a large number of cellular functions by binding with various partner proteins. The importance of its roles in many diseases requires an assay method to find regulatory compounds. However, the molecular characteristics of TCTP made it difficult to search for chemicals interacting with it. In this study, a tryptophan-based assay method was designed and Y151W mutant TCTP was constructed to search binding chemicals. Since there is no tryptophan in the native sequence of TCTP, the incorporation of tryptophan in the Y151W mutant was very effective to establish the method. A flavonoid library was employed to the assay with the method. With the native and Y151W mutant TCTPs, three flavonoids such as morin, myricetin and isobavachalcone have been found to interact with TCTP. Combined with native gel electrophoresis, the binding region of isobavachalcone was suggested to be the flexible loop of TCTP. This approach can be easily applicable to find binding compounds of proteins with similar molecular characteristics of TCTP.
Keywords
Transcriptionally controlled tumor protein (TCTP); Tryptophan; Florescence; Flavonoid; Native gel electrophoresis
Subject
Biology and Life Sciences, Biochemistry and Molecular Biology
Copyright:
This is an open access article distributed under the Creative Commons Attribution License which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
